LOCUS       sequence-243529-        5749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAGIC donor plasmid (Himar transposon with beta-lactamase/sfGFP 
            payload).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5749)
  AUTHORS   Ronda C, Chen SP, Cabral V, Yaung SJ, Wang HH
  TITLE     Metagenomic engineering of the mammalian gut microbiome in situ.
  JOURNAL   Nat Methods. 2019 Feb;16(2):167-170. doi: 10.1038/s41592-018-0301-y.
            Epub 2019 Jan 14.
  PUBMED    30643213
REFERENCE   2  (bases 1 to 5749)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41592-018-0301-y"; journalName: "Nat Methods"; date: 
            "2019-02"; volume: "16"; issue: "2"; pages: "167-170"
COMMENT     Sequence Label: pGT-Ah2
FEATURES             Location/Qualifiers
     source          1..5749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..69,70..861)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(219..238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             898..1611
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGFVTILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1671..1757
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1849..1876
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      3246..3791
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     3692..3711
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3908..3925
                     /label=L4440
                     /note="L4440 vector, forward primer"
     oriT            4061..4170
                     /note="incP origin of transfer"
     CDS             complement(4491..5150)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5065..5084
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5151..5255)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5323..5341
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
       61 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      121 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      181 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      241 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      301 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      361 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
      421 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
      481 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
      541 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
      601 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
      661 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
      721 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
      781 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
      841 tcactgatta agcattggta aattatattt aaactcaatt taaagtaggg agaatagatg
      901 cgtaaaggcg aagagctgtt cactggtttc gtcactattc tggtggaact ggatggtgat
      961 gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa
     1021 ctgacgctga agttcatctg tactactggt aaactgccgg taccttggcc gactctggta
     1081 acgacgctga cttatggtgt tcagtgcttt gctcgttatc cggaccacat gaagcagcat
     1141 gacttcttca agtccgccat gccggaaggc tatgtgcagg aacgcacgat ttcctttaag
     1201 gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac
     1261 cgcattgagc tgaaaggcat tgactttaaa gaagacggca atatcctggg ccataagctg
     1321 gaatacaatt ttaacagcca caatgtttac atcaccgccg ataaacaaaa aaatggcatt
     1381 aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca gcgtgcagct ggctgatcac
     1441 taccagcaaa acactccaat cggtgatggt cctgttctgc tgccagacaa tcactatctg
     1501 agcacgcaaa gcgttctgtc taaagatccg aacgagaaac gcgatcacat ggttctgctg
     1561 gagttcgtaa ccgcagcggg catcacgcat ggtatggatg aactgtacaa ataaaaacaa
     1621 taggccacat gcaactgtct agaatgcgag agtagggaac tgccaggcat caaataaaac
     1681 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     1741 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
     1801 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc
     1861 tgacggatgg cctttttgcg tttctacctg cagggcgcgc caagcagaag acggcatacg
     1921 aagaccgggg acttatcatc caacctgtta ggatccgttt taaatgatga aaagaaatat
     1981 ttagggaaga ttgtttcgac gcgaattgtt gatctggaaa atgatcacct tatcggacaa
     2041 gctttaaaat aggaggatat aaaaatcata tggaaaaaaa ggaatttcgt gttttgataa
     2101 aatactgttt tctgaaggga aaaaatacag tggaagcaaa aacttggctt gataatgagt
     2161 ttccggactc tgccccaggg aaatcaacaa taattgattg gtatgcaaaa ttcaagcgtg
     2221 gtgaaatgag cacggaggac ggtgaacgca gtggacgccc gaaagaggtg gttaccgacg
     2281 aaaacatcaa aaaaatccac aaaatgattt tgaatgaccg taaaatgaag ttgatcgaga
     2341 tagcagaggc cttaaagata tcaaaggaac gtgttggtca tatcattcat caatatttgg
     2401 atatgcggaa gctctgtgcg aaatgggtgc cgcgcgagct cacatttgac caaaaacaac
     2461 gacgtgttga tgattctaag cggtgtttgc agctgttaac tcgtaataca cccgagtttt
     2521 tccgtcgata tgtgacaatg gatgaaacat ggctccatca ctacactcct gagtccaatc
     2581 gacagtcggc tgagtggaca gcgaccggtg aaccgtctcc gaagcgtgga aagactcaaa
     2641 agtccgctgg caaagtaatg gcctctgttt tttgggatgc gcatggaata atttttatcg
     2701 attatcttga gaagggaaaa accatcaaca gtgactatta tatggcgtta ttggagcgtt
     2761 tgaaggtcga aatcgcggca aaacggcccc acatgaagaa gaaaaaagtg ttgttccacc
     2821 aagacaacgc accgtgccac aagtcattga gaacgatggc aaaaattcat gaattgggct
     2881 tcgaattgct tccccacccg ccgtattctc cagatctggc ccccagcgac tttttcttgt
     2941 tctcagacct caaaaggatg ctcgcaggga aaaaatttgg ttgcaatgaa gaggtgatcg
     3001 ccgaaactga ggcctatttt gaggcaaaac cgaaggagta ctaccaaaat ggtatcaaaa
     3061 aattggaagg tcgttataat cgttgtatcg ctcttgaagg gaactatgtt gaataagcgg
     3121 ccgccaccgc ggtggagggg aattaacaac ttatatcgta tggggctgac ttcaggtgct
     3181 acatttgaag agataaattg cactgaaatc tagaaatatt ttatctgatt aataagatga
     3241 tcttcttgag atcgttttgg tctgcgcgta atctcttgct ctgaaaacga aaaaaccgcc
     3301 ttgcagggcg gtttttcgaa ggttctctga gctaccaact ctttgaaccg aggtaactgg
     3361 cttggaggag cgcagtcacc aaaacttgtc ctttcagttt agccttaacc ggcgcatgac
     3421 ttcaagacta actcctctaa atcaattacc agtggctgct gccagtggtg cttttgcatg
     3481 tctttccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cggactgaac
     3541 ggggggttcg tgcatacagt ccagcttgga gcgaactgcc tacccggaac tgagtgtcag
     3601 gcgtggaatg agacaaacgc ggccataaca gcggaatgac accggtaaac cgaaaggcag
     3661 gaacaggaga gcgcacgagg gagccgccag ggggaaacgc ctggtatctt tatagtcctg
     3721 tcgggtttcg ccaccactga tttgagcgtc agatttcgtg atgcttgtca ggggggcgga
     3781 gcctatggaa aaacggcttt gccgcggccc tctcacttcc ctgttaagta tcttcctggc
     3841 atcttccagg aaatctccgc cccgttcgta agccatttcc gctcgccgca gtcgaacgac
     3901 cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc cagatcctct acgccggacg
     3961 catcgtggcc ggatcaattc cgtgataggt gggctgccct tcctggttgg cttggtttca
     4021 tcagccatcc gcttgccctc atctgttacg ccggcggtag ccggccagcc tcgcagagca
     4081 ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc
     4141 gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc gttggataca ccaaggaaag
     4201 tctacacgaa ccctttggca aaatcctgta tatcgtgcga aaaaggatgg atataccgaa
     4261 aaaatcgcta taatgacccc gaagcagggt tatgcagcgg aaaacggaat tgatccccct
     4321 cgaattgacg cgtaaccagc aatagacata agcggctatt taacgaccct gccctgaacc
     4381 gacgaccggg tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt
     4441 caggcgtagc aaccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg
     4501 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
     4561 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     4621 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
     4681 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
     4741 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
     4801 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
     4861 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
     4921 attgccatac gtaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
     4981 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
     5041 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
     5101 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat actcttcctt
     5161 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5221 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5281 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5341 ccctttcgtc acgcgtcctc ggtacctaac aggttggatg ataagtcccc ggtcttcgta
     5401 tgccgtcttc tgcttggcgc gccctcgagc aattgccgac cgaattttta tgtcgtaaag
     5461 aggggctttg cagggggtgg actcagaaag atgagaatag atgactattg tagttgaaac
     5521 acatagaaag ttgctgatat acagaccgat acgcatatcg ggatgaacca tgagtacgtt
     5581 cttttctcaa aaaacataaa tattcgaaaa gagatgcaat aaattaagga gaggttatag
     5641 ttttaaatga tgaaaagaaa tatttaggga agattgtttc gacgcgaatt gttgatctgg
     5701 aaaatgatca ccttatcgga caagctttaa aataggagga tataaaaat
//
